In vitro test

Global Leader of Human Skin & Bio Research

17

Wound healing assay

The effect of cosmetic ingredients and natural substances on cell migration is evaluated by “Wound gap” in a cell monolayer. The healing of this gap by cell migration closes the wound, and the images are compared to quantify the migration rate of the cells.

Duration

8 weeks

16

Cytotoxicity Test

The cytotoxicity of the cosmetics ingredients and natural substances is determined by MTT, XTT, WST-1 and Alamar blue assay

Duration

6 weeks

15

Collagen synthesis assay

The effect of cosmetics ingredients and natural substances on collagen synthesis is evaluated in human dermal fibroblast with procollagen biosynthesis with Procollagen Type I C-peptide EIA kit in accordance with the Ministry of Food and Drug Safety guideline for evaluation of the effectiveness of functional cosmetics?intercellular collagen synthesis assay

Duration

10 weeks

14

Collagenase inhibition assay

The effect of cosmetics ingredients and natural substances on collagenase inhibition is evaluated in cells induced collagenase by ELISA assay in accordance with the Ministry of Food and Drug Safety guideline for evaluation of the effectiveness of functional cosmetics-Intracellular collagenase activity inhibition test

Duration

10 weeks

13

Elastase inhibition assay

The effect of cosmetics ingredients and natural substances on elastase inhibition is determined by enzyme assay using elastase from human dermal fibroblast cells in accordance with the Ministry of Food and Drug Safety guideline for evaluation of the effectiveness of functional cosmetics- elastase inhibition assay

Duration

4 weeks

12

Tyrosinase inhibition assay

The effect of cosmetics ingredients and natural substances on tyrosinase activity is evaluated by in vitro enzyme assay using mushroom tyrosinase and L-tyrosine as a substrate in accordance with the Ministry of Food and Drug Safety guideline for evaluation of the effectiveness of functional cosmetics- In vitro tyrosinase inhibition test

Duration

4 weeks

11

DOPA oxidation inhibition assay

The inhibitory effect of cosmetics ingredients and natural substances on DOPA oxidation is evaluated by in vitro enzyme assay using mushroom tyrosinase and L-DOPA as a substrate in accordance with the Ministry of Food and Drug Safety guideline for evaluation of the effectiveness of functional cosmetics- In vitro DOPA oxidation inhibition test

Duration

4 weeks

10

Melanin synthesis inhibition assay

The effect of cosmetics ingredients and natural substances on melanin synthesis inhibition is determined in B16F10 cells stimulated with melanin elevating agents in accordance with the Ministry of Food and Drug Safety guideline for evaluation of the effectiveness of functional cosmetics-melanin synthesis inhibition test

Duration

10 weeks

9

Anti-oxidant assay (DPPH)

The effect of cosmetics ingredients and natural substances on anti-oxidant activity is determined by the DPPH radical scavenging method

Duration

4 weeks

8

Oxygen Radical Absorbance Capacity assay (ORAC)

Oxygen radical absorbance capacity (ORAC) is a method of measuring antioxidant capacities in cosmetics ingredients and natural substances in vitro. The effect of sample on anti-oxidant activity is determined by the ROO? (peroxyl radical) scavenging method

Duration

6 weeks

7

Total polyphenol content assay

Total polyphenol content is determined using a spectrophotometric assay based on the reduction of Folin-Ciocalteau reagent by phenolic compounds, with a concomitant formation of a blue complex

Duration

4 weeks

6

Nitrogen oxide (NO) assay

The inhibitory effect of cosmetics ingredients and natural substances on NO production is evaluated in LPS-induced Raw264.7 cells

Duration

8 weeks

5

Western blot

The effect of cosmetics ingredients and natural substances on the specific protein related to inflammation, skin aging, skin barrier and hydration is evaluated by western blot, an analytical technique used to detect the specific protein from a cell extract

Efficacy

Factor (cytokine)

Anti-inflammation

IL-1α, IL-1β, IL-6, IL-8, TNF-α, COX-2

Anti-aging

Collagen type I

MMP-1 (collagenase)

MMP-2 (gelatinase)

MMP-3 (Stromelysin)

MMP-9 (gelatinase)

TIMP-1 (inhibitor of the MMPs)

Skin barrier & hydration

Involucrin, Filaggrin, AQP-3, HAS-2, Loricrin, Keratin

Duration

12 weeks

4

RT-PCR & RT-qPCR

The effect of cosmetics ingredients and natural substances on mRNA level of genes related to inflammation, skin aging, skin barrier and hydration is evaluated using RT-PCR & RT-qPCR

Duration

12 weeks

3

ELISA assay

The effect of cosmetics ingredients and natural substances on the specific inflammatory cytokines related to inflammation is evaluated by ELISA assay

Duration

10 weeks

2

Alternative test using artificial skin (non-GLP)

Skin initiaion of the test material is evaluated by skin irritation test using reconstructed human epidermis in accordance with the OECD guideline (TG 439) in Ministry of Food and Drug Safety guideline

Duration

12 weeks

1

3T3 NRU Phototoxicity Test

The effect of cosmetics ingredients and natural substances on phototoxicity is evaluated using 3T3 NRU assay in accordance with the Ministry of Food and Drug Safety guideline for in vitro 3T3 NRU phototoxicity Test